P 69 ISSRNS 2012: Abstracts / Synchrotron Radiation in Natural Science Vol. 11, No 1 – 2 (2012)
SR-FTIR SPECTROSCOPY IN STUDY OF THE DOUBLE STAND BREAKS IN SINGLE CELLS IRRADIATED BY PROTON MICROBEAM
A. Wieche´c1∗, E. Lipiec1, J. Lekki1, M. Wide l2, and W.M. Kwiatek1
1The Henryk Niewodniczanski Institute of Nuclear Physics, Polish Academy of Sciences, Krakow, Poland
2Silesian University of Technology, Department of Automatics, Electronics and Informatics, Gliwice, Poland
Keywords: synchrotron radiation, double strand breaks, single cell irradiation, proton microbeam
∗e-mail : Anna.Wiechec@ifj.edu.pl
The double strand breaks (DSBs) in DNA are the critical cell damage if unrepaired they may lead to cell death. Recently, many published data indi- cate that the formation of DSBs in DNA is followed by the rapid local phosphorylation of histone H2AX.
The formation of discrete nuclear foci called γH2AX foci is observed [1, 2]. The γH2AX foci, a sensitive marker of DNA double strand breaks, might be vi- sualized after conjugation with anti-γH2AX mono- clonal antibodies joint with fluorochrome.
From the possible physical methods, the SR- FTIR microspectroscopy is well known for its uniqueness as a noninvasive tool in identifying vi- brational structure of biological materials. It has become a potential analytical method in single cells studies [3, 4].
The aim of this study was the evaluation of the DNA double strand breaks in proton irradiated sin- gle cells with application of SR-FTIR spectroscopy.
The study tried to answer the question if and how the chemicals used in γH2AX test affects the SR- FTIR cell spectra.
The DNA damage in single cells from prostate cancer PC3 cell line were inducted with proton mi- crobeam (2 MeV energy; 50 and 2000 protons per cell). The evaluation of DNA double strand breaks was done in γH2AX test and by means of SR-FTIR spectroscopy. For the irradiation treatment the cells were seeded on the silicon nitride windows.
Detailed description of γH2AX test is presented in [1, 2]. The 100 nucleus of irradiated and control cells were examined under fluorescent microscope Zeiss Axio Imager Z1.
SR-FTIR cell spectra were analyzed in transmis- sion mode with a resolution of 4 cm−1 in spectral
range 400 cm−1 – 4000 cm−1.
The application of both biochemical and physi- cal methods enabled detection of changes in irradi- ated cells at the chemical bonds level. The compar- ison of the results obtained from both methods is discussed.
Acknowledgments: This work was supported by the European Community’s Seventh Framework Pro- gramme (FP7/2007 – 2013) under grant agreement n◦ 226716. The data were obtained during the realization of PSI Synchrotron project 20110190.
References
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